Infant Formula With a Specific Blend of Five Human Milk Oligosaccharides Drives the Gut Microbiota Development and Improves Gut Maturation Markers: A Randomized Controlled Trial

Abstract

BackgroundHuman milk oligosaccharides (HMOs) have important biological functions for a healthy development in early life.ObjectiveThis study aimed to investigate gut maturation effects of an infant formula containing five HMOs (2′-fucosyllactose, 2′,3-di-fucosyllactose, lacto-N-tetraose, 3′-sialyllactose, and 6′-sialyllactose).MethodsIn a multicenter study, healthy infants (7–21 days old) were randomly assigned to a standard cow’s milk-based infant formula (control group, CG); the same formula with 1.5 g/L HMOs (test group 1, TG1); or with 2.5 g/L HMOs (test group 2, TG2). A human milk-fed group (HMG) was enrolled as a reference. Fecal samples collected at baseline (n∼150/formula group; HMG n = 60), age 3 (n∼140/formula group; HMG n = 65) and 6 (n∼115/formula group; HMG n = 60) months were analyzed for microbiome (shotgun metagenomics), metabolism, and biomarkers.ResultsAt both post-baseline visits, weighted UniFrac analysis indicated different microbiota compositions in the two test groups (TGs) compared to CG (P < 0.01) with coordinates closer to that of HMG. The relative abundance of Bifidobacterium longum subsp. infantis (B. infantis) was higher in TGs vs. CG (P < 0.05; except at 6 months: TG2 vs. CG P = 0.083). Bifidobacterium abundance was higher by ∼45% in TGs vs. CG at 6-month approaching HMG. At both post-baseline visits, toxigenic Clostridioides difficile abundance was 75–85% lower in TGs vs. CG (P < 0.05) and comparable with HMG. Fecal pH was significantly lower in TGs vs. CG, and the overall organic acid profile was different in TGs vs. CG, approaching HMG. At 3 months, TGs (vs. CG) had higher secretory immunoglobulin A (sIgA) and lower alpha-1-antitrypsin (P < 0.05). At 6 months, sIgA in TG2 vs. CG remained higher (P < 0.05), and calprotectin was lower in TG1 (P < 0.05) vs. CG.ConclusionInfant formula with a specific blend of five HMOs supports the development of the intestinal immune system and gut barrier function and shifts the gut microbiome closer to that of breastfed infants with higher bifidobacteria, particularly B. infantis, and lower toxigenic Clostridioides difficile.Clinical Trial Registration[https://clinicaltrials.gov/ct2/show/], identifier [NCT03722550].

Functional effects of human milk oligosaccharides (HMOs)

“…Three major HMO categories are present in human milk of secretor mothers 6 , 14 , 15 , 25 , 26 Neutral fucosylated HMOs (35–50%; e.g., 2′-FL and DFL) Acidic sialylated HMOs (12–14%) e.g., 3′-SL and 6′-SL Neutral non-fucosylated HMOs (42–55%, e.g., LNnT, LNT). …”

“… Test group and breastfed infants had significant lower fecal calcium excretion ( p < 0.005) and fecal pH ( p < 0.001) and higher lactate ( p < 0.001) compared to standard formula group. Bosheva et al 25 2022 2’-FL, DFL, LNT, 3’-SL, 6’-SL Bulgaria, Hungary, and Poland 2018 and November 2021 Healthy and full-term, with birth weight between 2,500 and 4,500 g, and aged≥7– ≤21 days at enrollment. N = 50 Test formula 1: 1.5 g/L of a mix of 2’-FL, DFL, LNT, 3’-SL, 6’-SL Test formula 2: 2.5 g/L of a mix of 2’-FL, DFL, LNT, 3’-SL, 6’-SL Standard cow’s milk-based formula Breastfed infants 15 months Microbiota in the two test groups were significantly ( p < 0.01) different in comparison to the standard cow’s milk formula group (SFG).…”

“…However, since there were only six infants no conclusions can be drawn 160 . Another multicenter study examined fecal cultures of infants receiving either a formula with a mix of five HMOs at a concentration of 1.5 g/L, a formula with a mix of five HMOs at a concentration of 2.5 g/L, a non-supplemented formula, or breast milk 25 . The microbiota composition of infants receiving formulas supplemented with HMOs was significantly different to those in the non-supplemented group and were closer to the composition of the breastfed infants.…”

Effects of a Novel Infant Formula on Weight Gain, Body Composition, Safety and Tolerability to Infants: The INNOVA 2020 Study

“…The main objective of the study was to evaluate whether the weight gain was equivalent between treatment groups receiving formula 1 and 2, and it was decided that the sample size of the trial would be 210 children (70/group) based on the primary variable weight gain, following the “Guidelines from the American Academy of Pediatrics” issued by the Task Force on Clinical Testing of Infant Formulas of the American Academy of Pediatrics [ 44 ]. Previous studies carried out on children fed from 0 to 6 months with different formulations of infant formula have shown a mean weight gain of around 20–25 g/day with a standard deviation between 5 and 6 g/day [ 46 ]. In most of these studies, a difference in mean weight gain of 3 g/day was considered clinically relevant.…”

Effects of a novel infant formula on weight gain and body composition of infants: The INNOVA 2020 study

“…American Academy of Pediatrics Committee on Nutrition et al, 1988). Previous studies carried out on children fed from 0 to 6 months with different formulations of infant formula have shown a mean weight gain of around 20-25 g/day with a standard deviation between 5 and 6 g/day (Bosheva et al, 2022). In most of these studies, a difference in mean weight gain of 3 g/day was considered clinically relevant.…”